Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6506604

Title:Method for production of high titer virus and high efficiency retroviral mediated transduction of mammalian cells
Inventors:Finer, Mitchell H.; San Carlos, CA, USA
Dull, Thomas J.; San Francisco, CA, USA
Zsebo, Krisztina M.; Woodside, CA, USA
Cooke, Keegan; Palo Alto, CA, USA
Farson, Deborah A.; Oakland, CA, USA
Summary:This invention introduces a high efficiency retroviral packaging plasmid system for producing virions which may be used to transduce human cells such as hematopoietic stem cells, T or B cells, with foreign genes. The invention relates to a novel retroviral packaging system in which retroviral packaging plasmids and packagable vector transcripts are produced from high expression plasmids after stable or transient transfection in mammalian cells. Further disclosed are methods by which high titers of a recombinant retrovirus are produced in these transfected mammalian cells which can then transduce a mammalian target cell by cocultivation or supernatant infection involving T cells and human hematopoietic stem cells. Applications are included for the rapid production of high titer viral supernatants, and for improved transduction with high efficiency cells that are refractory to transduction by conventional means.
Abstract:The invention provides a novel retroviral packaging system, in which retroviral packaging plasmids and packagable vector transcripts are produced from high expression plasmids after stable or transient transfection in mammalian cells. High titers of recombinant retrovirus are produced in these transfected mammalian cells and can then transduce a mammalian target cell by cocultivation or supernatant infection. The methods of the invention include the use of the novel retroviral packaging plasmids and vectors to transduce primary human cells, including T cells and, human hematopoietic stem cells, with foreign genes by cocultivation or supernatant infection at high efficiencies. The invention is is useful for the rapid production of high titer viral supernatants, and to transduce with high efficiency cells that are refractory to transduction by conventional means.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6506604
Application Number:US2001000944411
Date Filed:04/09/2001
Date Published:14/01/2003
Assignee:Cell Genesys, Inc., Foster City, CA, USA


 
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